26 results about "Mgb probe" patented technology

The invention discloses a composition, a kit and a method for detecting a mitochondrial gene of sea eel. A primer probe composition targeting the D-LOOP gene of the sea eel mitochondria is designed, and the nucleotide sequences of the primer probe composition correspond to SEQ ID No.1-3. On the basis, the invention further constructs and optimizes a specific real-time fluorescent PCR detection method for the D-LOOP gene of the sea eel mitochondria. According to the method, a part of sequence of the D-LOOP gene of the mitochondria of the sea eel is taken as a target region, a primer and a TaqMan-MGB probe are designed in a targeted manner, and finally a set of rapid, sensitive and high-specificity detection system is established. The detection system can be effectively applied to adulteration and germplasm identification work of the sea eel, and has a very wide industrial application prospect.

The invention relates to the technical field of pathogenic bacterium detection, in particular to a method and a kit for detecting helicobacter pylori gene mutation in excrement. According to the technical scheme, the kit for detecting helicobacter pylori gene mutation in excrement is developed by combining multiple advanced technical means such as ARMS-PCR design, blocking primers, locked nucleic acid modification (peptide nucleic acid modification), MGB probes, nested PCR design and degenerate base design. The kit can detect nine gene mutation sites related to the drug resistance of helicobacter pylori at one time, and has a wide coverage range. The kit in the scheme can be used for detecting the drug resistance of quinolone antibiotics and / or clarithromycin antibiotics of helicobacter pylori, and the technical problem that a high-specificity and high-sensitivity excrement helicobacter pylori gene mutation detection method is lacked in the prior art is solved. The method is simple and convenient to operate and can be completed within 1-2 hours, the detection cost is reduced, and the method has ideal application and popularization prospects.

The invention provides a primer probe, a kit and a method for identifying A1 / A2-beta casein in milk and dairy products. The primer probe comprises an allele specific primer A1A2-F, an allele specific primer A1A2-R, a probe A2-P and a probe A1-P. The invention further discloses a kit for detecting the allele specific primer A1A2-F, the allele specific primer A1A2-R and the probe A1-P. According to the present invention, based on the 67th single nucleotide polymorphism (SNP) of the ruminant A1 / A2-beta casein gene (CSN2), the gene typing method with characteristics of high specificity and high sensitivity is constructed; according to the method, an ARMS-PCR technology, a locked nucleic acid (LNA) modification strategy and an MGB probe signal detection system are integrated, so that the A1 / A2 allele is accurately distinguished.

Taqman-MGB probe real-time fluorescence quantitative PCR detection system, kit, method and application of Vibrio mimics, the present invention uses three O antigen serotypes of Vibrio mimics wzy Using the gene as the target gene, Taqman primers and MGB probes specific for each serotype were designed and screened, and a real-time fluorescence PCR detection method was established. The detection system provided by the present invention can accurately, quickly and sensitively perform molecular biological detection on the three Vibrio mimics.

The present invention discloses a primer and probe combination and a kit for detecting galactose-1-phosphate uridyltransferase gene polymorphism, which relates to the technical field of in vitro detection. The primer and probe combination for detecting galactose-1-phosphate uridyltransferase is detected in two categories, and each category of primer and probe group has 3 primer and probe combinations, which can simultaneously detect 8 mutation sites of the galactose-1-phosphate uridyltransferase gene, shortening the diagnosis time and reducing the detection cost; Taqman probes and MGB probes are used in the same category of primer and probe group, which can better distinguish the Tm values of heterozygous samples, reduce the problems of missed detection or genotype misjudgment, and improve the detection sensitivity and accuracy; In addition, the β-actin gene internal reference is used for quality control, improving the comparability of the detection.

The application discloses primers and a probe for real-time fluorescent quantitative PCR detection of a porcine epidemic diarrhea virus G2c subtype and application thereof and belongs to the field of animal virus molecular biology testing methods. The application compares S genes of different gene subtype strains of the porcine epidemic diarrhea virus, designs a pair of primers and a Taqman-MGB probe on the basis of a mutation site specific to the G2c subtype strain, and proves that the primers and the probe can effectively distinguish the G2c and other PEDV gene subtype strains through virus liquids and clinical samples of different gene subtype strains of the PEDV, meanwhile, the primers and the probe have no cross reaction to other viruses of the porcine enteric diarrhea syndrome, and the detection sensitivity of the kit can reach 5 copies / muL. The application can realize specific and sensitive detection of the G2c subtype, and has a positive significance for diagnosis and epidemiological investigation of the porcine epidemic diarrhea virus.

The invention discloses a primer probe set for identifying coreius guichenoti and application. The primer probe group comprises an upstream primer COX1-F1 (SEQ ID NO. 1), a downstream primer COX1-R1 (SEQ ID NO. 2), and a TaqMan-MGB probe COX1-P (SEQ ID NO. 3), and is characterized in that the upstream primer COX1-F1 (SEQ ID NO. 1), the downstream primer COX1-R1 (SEQ ID NO. 2) and the TaqMan-MGB probe COX1-P (SEQ ID NO. Experiments prove that the primer probe group has extremely high specificity on coreius guichenoti, and has no cross reaction with other common fishes which belong to the same genus and are distributed in the same watershed; the qPCR detection method is high in sensitivity, the lowest detection limit can reach 10 copies / mu L, and the linearity of a standard curve is excellent (Rgt; the invention provides a high-sensitivity and high-specificity molecular detection tool and a related detection method for rapid species identification, resource monitoring and endangered mechanism research of coreius guichenoti, and particularly has a wide application prospect in the aspect of non-invasive environmental DNA monitoring.

The application belongs to the technical field of biological detection, and particularly relates to a SNP site for distinguishing between MS-H vaccine strains and wild strains of synovial bursa mycoplasma, a kit and application. ktrb The application first designs specific primers and MGB probes for specific conservative gene sequences, and establishes a double qPCR kit for distinguishing between MS-H vaccine strains and wild strains. Experiments in vitro prove that the kit is simple to operate, high in sensitivity, strong in specificity, and good in repeatability. The gene site and the detection kit have the advantages of avoiding the deficiency that the commonly used genes are prone to revert mutation, can be used for clinical rapid diagnosis of MS vaccine strains and wild strains, improve the accuracy of detection, promote population purification, and have certain commercial value.

The invention discloses a detection method for noninvasive prenatal detection of fetal trisomy 21 syndrome based on digital PCR, and relates to the technical field of digital PCR. According to the invention, a target area detection amplification primer and a Taqman-MGB probe are designed according to different sequences of human chromosome 21 genes, and a specific common amplification primer and a specific Taqman-MGB probe are designed according to similar sequences of the human chromosome 21 genes. Accurate detection of a low-proportion trisomy 21 syndrome positive sample is realized, and the omission ratio is greatly reduced. Compared with next-generation sequencing, multiple qPCR technologies and the like, the detection method provided by the invention has the advantages of simplicity in operation, simplicity in data analysis, short detection period, low cost and the like, meanwhile, by innovatively using a similar sequence design technology, high consistency of amplification efficiency of a detection gene and a reference gene is ensured, and the detection accuracy is improved. The detection sensitivity and accuracy are obviously improved.

The invention relates to the technical field of nucleic acid molecule detection, and discloses a fusion gene detection system based on multiple fluorescent quantitative PCR. The TaqMan-MGB probe group, the specific primer pair and the PCR buffer system are optimally designed, so that the binding specificity of the probe group and the primer pair is effectively improved, the binding efficiency is improved, and the sensitivity of fusion gene detection is improved. Specifically, the probe is arranged across the fusion connection area, so that the specificity is ensured to the maximum extent. The primer crosses two sides of a fusion breakpoint to ensure that only fused templates can be amplified at the same time. The multiple fluorescent quantitative PCR detection system provided by the invention can simultaneously amplify and monitor the existence condition of various thyroid cancer fusion genes in one tube reaction.

The invention discloses a specific primer probe mixture and a kit for BKV virus detection. The specific primer probe mixture for detecting the BKV virus comprises a forward primer as shown in SEQ ID NO: 1, a reverse primer as shown in SEQ ID NO: 2 and an MGB probe as shown in SEQ ID NO: 3. According to the present invention, the unique target spot selection is provided, the specific nucleotide fragment (4673-4722bp) of the BKV VP1 gene is selected as the detection target sequence, and the MGB probe is adopted, such that the annealing temperature and the binding specificity are improved, and the non-specific amplification is effectively inhibited.

The invention relates to a primer and a TaqMan-MGB probe group for fluorescent quantitative PCR (Polymerase Chain Reaction) detection of virulent and attenuated strains of classical waterfowl parvovirus, the primer and the TaqMan-MGB probe group comprise a fluorescent quantitative PCR detection primer and a probe group for virulent and attenuated strains of goose parvovirus respectively, and the sequences of the primer and the probe group are shown as SEQ ID NO.1-5 respectively; the primer and the TaqMan-MGB probe group further comprise a fluorescent quantitative PCR detection primer and a probe group for virulent and attenuated muscovy duck parvovirus respectively, and the sequences of the primer and the probe group are shown as SEQ ID NO.6-9 respectively. According to the invention, through systematic test verification and reaction condition optimization, an optimal primer-probe combination is screened out, a GPV TaqMan-MGB dual fluorescent quantitative PCR detection method and an MDPV TaqMan dual fluorescent quantitative PCR detection method are respectively and successfully established, and the detection of virulent and attenuated classical waterfowl parvovirus is realized.

The application belongs to the technical field of microorganism detection, and particularly relates to detection primers and probes for Euryphallus sinensis and application thereof. The application establishes a real-time fluorescent quantitative PCR method for the pathogen based on TaqMan-MGB probes according to the small subunit ribosomal RNA (SSUrRNA) gene of Euryphallus sinensis, and designs detection primers based on isothermal amplification technology. With the constructed recombinant plasmid standard as a template, the detection sensitivity of the quantitative PCR detection method for the target gene is 9.58 copies / reaction. The detection primers for isothermal amplification can be detected under simple constant temperature conditions, and the color change of the detection product after adding nucleic acid dye can realize on-site rapid diagnosis of the pathogen.

Described herein are compositions, methods, and systems for the detection and quantification of 5-plex and 6-plex multiplex (5 or 6 amplified nucleic acid targets) with QSY2™ probes. Some embodiments relate to detection and quantification of 5-plex and 6-plex multiplex (5 or 6 amplified nucleic acid targets) with a combination of QSY2™ probes and QSY™ probes, or a combination of QSY2™ probes with MGB probes, in a single reaction. Other embodiments relate to detection and quantification of 5-plex and 6-plex multiplex (5 or 6 amplified nucleic acid targets) with a combination of QSY2™ probes with QSY™ probes, or a combination of QSY2™ probes and MGB probes in a single reaction.

The invention discloses a screening method and application of blast-resistant rice, relates to the technical field of nucleotide molecule screening, and in particular relates to a screening method of blast-resistant rice and a TaqMan-MGB probe for specifically detecting a blast-resistant gene Pigm of rice. The method can be used for performing gene molecule on a plurality of rice samples in a high-throughput and high-efficiency manner and automatically screening out the rice with the homozygous Pigm genotype, so that the effect of quickly screening the rice blast-resistant rice is realized.

The application relates to the technical field of molecular biology detection, and discloses a z. laticeps environmental DNA detection kit, which comprises a primer probe composition for real-time fluorescent PCR detection designed based on a z. laticeps mitochondrial Cytb gene, the composition comprises an upstream primer, a downstream primer and a TaqMan-MGB probe, the nucleotide sequence of the upstream primer is shown in SEQ ID No. 1, the nucleotide sequence of the downstream primer is shown in SEQ ID No. 2, and the nucleotide sequence of the TaqMan-MGB probe is shown in SEQ ID No. 3; the 5' end of the TaqMan-MGB probe is modified with a fluorescent reporter group, and the 3' end is coupled with a small groove binder and a non-fluorescent quenching group; the application creates a special primer probe composition and a complete detection system for z. laticeps environmental DNA detection, realizes high specificity and high sensitivity detection of z. laticeps, and uses z. laticeps specific mitochondrial genes as detection targets; the TaqMan-MGB probe can accurately identify slight differences in gene sequences and effectively distinguish z. laticeps from closely related species in the same region.

The invention discloses a molecular marker related to drug resistance of Brucella melitensis compound sulfamethoxazole and application of the molecular marker, and relates to the technical field of biological detection. The molecular marker comprises an SNP (Single Nucleotide Polymorphism) site; the SNP site is located at the 877673 site of a Brucella melitensis chromosome 1, and is G / A mutation. A specific qPCR primer pair and an MGB probe pair are designed on the basis of the molecular marker and are used for specifically detecting rough variation of Brucella melitensis and drug resistance of compound sulfamethoxazole, and experimental operations such as pathogen separation and rough phenotype identification are not needed; the kit can be used for rapidly and intuitively identifying whether a sample contains rough variation and compound sulfamethoxazole-resistant brucella melitensis, can be applied to rapid detection of rough variation and drug-resistant bacteria in clinical brucellosis and abortion regulation processes, and is beneficial to promoting the brucellosis prevention and control process.

The application belongs to the technical field of molecular biology and blood grouping, and particularly relates to a Taqman-MGB probe-based fluorescent quantitative PCR detection primer group and kit and application thereof in MNS blood group system genotyping. The application designs specific primers based on the MGB probe enhanced single base mismatch recognition ability, generates specific amplification signals for specific sites of the MNS blood group system, and has a detection sensitivity as low as 0.1 ng genomic DNA, strong specificity and high sensitivity. The application optimizes the proportion of Probe 1 and Probe 2 probes in the reaction system, effectively balances the signal intensity of different fluorescence channels, makes the target and non-target channel CT values of the homozygote sample significantly different (usually greater than or equal to 5), and improves the objectivity and repeatability of interpretation. The method of the application is simple, rapid and efficient, does not need subsequent steps such as electrophoresis and sequencing, and is suitable for high-throughput genotyping detection of clinical and laboratory blood groups.

The invention discloses a composition, a kit and a method for detecting Huso dauricus mitochondrial genes. A specific primer probe composition is designed aiming at the Huso dauricus mitochondrial ND4 gene, and the corresponding nucleotide sequences of the specific primer probe composition are SEQ ID No.1-3. By taking the Huso dauricus mitochondrial ND4 gene as a core technical support, the invention further constructs and optimizes a real-time fluorescent PCR specific detection scheme based on the Huso dauricus mitochondrial ND4 gene. According to the scheme, a part of sequence of a Huso dauricus mitochondrial ND4 gene is selected as a target fragment, a primer and a TaqMan-MGB probe are directionally designed, and finally a set of detection system with rapidness, high sensitivity and strong specificity is successfully established. The detection system can be effectively applied to adulteration identification and germplasm resource identification of huso dauricus products, and has very wide industrial popularization and application prospects.

The present invention belongs to the field of biochemistry, and specifically relates to a Taq enzyme mutant and its application in SNP-related research. The specific technical solution is as follows: a Taq enzyme mutant, which is mutated from wild-type C-Taq enzyme, and the amino acid sequence of wild-type C-Taq enzyme is shown in SEQ ID NO: 1. Based on the wild-type C-Taq enzyme, at least the following mutations are carried out: the threonine at position 506 of the wild-type C-Taq enzyme is mutated to lysine. The present invention analyzes the key active sites of wild-type C-Taq enzyme and performs site-directed mutagenesis, and uses multiple pairs of SNP genotyping primers for high and low GC target fragments to test the candidate mutant enzymes, and screens two enzymes with better discrimination ability for single-base mismatches. The single-base mismatch probe of the mutant enzyme provided by the present invention has a lower peak and higher genotyping accuracy. The mutant enzymes provided by the present invention can amplify well at high, medium and low GC sites, with wide universality; using these mutant enzymes can effectively save the cost of screening MGB probes.

The invention discloses a detection kit for S gene mutation of feline infectious peritonitis virus and a use method. A primer for detecting S gene mutation adopts ARMS-PCR combined with an LNA technology, a mutation primer is designed aiming at a mutation basic group, and the mutation primer is completely matched with a mutation sequence and is mismatched with the 3'tail end of a wild type template; the 3'terminal basic group of the mutation primer is modified by LNA, and the specificity level of the primer is improved, so that the mutation primer only amplifies the corresponding mutation type template, and if the mutation primer is combined with the wild type template, the amplification efficiency is extremely low or the amplification cannot be realized, thereby realizing the detection of the low-content mutation sequence under the background of the high wild type gene. Due to the fact that the mutation rate of the base in the S region is high, the Taqman-MGB probe is adopted, the length of the probe sequence is shortened, the specificity of the Taqman-MGB probe is increased, hypoxanthine modification is conducted on the base with high SNP in the probe sequence, mutation of the base with the high SNP in the probe sequence is reduced, and missing detection is avoided. The ARMS-PCR technology is combined with the Taq-MGB real-time fluorescence PCR method, so that the result can be judged according to a real-time fluorescence amplification curve, and subsequent operation is not needed.

The present application relates to the technical field of nucleotide molecule screening, in particular to a mutagenesis screening method and application of a rice herbicide-resistant genotype. The present application designs a qPCR reaction system based on a specific SNP site region of a mutant ALS gene, realizes the effect of high-throughput and large-scale screening of a rice parent containing a mutant ALS gene, and can significantly improve the efficiency of rice screening and breeding. The present application designs a TaqMan-MGB probe based on a specific SNP site region of a mutant ALS gene, which can form a hybrid molecule with a specific SNP site with high specificity and high accuracy, ensuring the accuracy and reproducibility of qPCR.

The invention discloses a primer, a probe, an implementation method and a detection system for qPCR detection of SQLE drug-resistant mutation of dermatophytes, and the method comprises the following steps: identifying a plurality of key mutation sites related to drug resistance by comparing an obtained dermatophytes SQLE gene sequence with a reference sequence in an NCBI database; designing a specific primer, a TaqMan-MGB fluorescent probe and a wild type closed probe modified by the MGB probe based on the mutation site, and verifying the sensitivity and the specificity through the strain level; the method comprises the following steps: constructing a mixed DNA sample model containing drug-resistant strain DNA, wild type strain DNA and human DNA, optimizing a reaction system of single and multiple qPCR detection, and determining an optimal reaction system of qPCR detection. According to the specific primer and probe for qPCR detection of dermatophyte SQLE drug-resistant mutation with specificity meeting requirements and an optimized reaction system for qPCR detection, whether dermatophyte SQLE drug-resistant mutation exists or not can be quickly verified, the sensitivity and specificity of dermatophyte drug-resistant detection are improved, early precise treatment is facilitated, and prognosis of patients is improved.

The invention discloses a Taqman-MGB probe real-time fluorescent quantitative PCR (polymerase chain reaction) detection system, a kit, a method and application of vibrio mimicus and discloses a Taqman-MGB probe real-time fluorescent quantitative PCR detection system, a kit, a method and application of the Taqman-MGB probe real-time fluorescent quantitative PCR detection system and the kit. Wzy genes of three O antigen serotypes of vibrio mimicus are used as target genes, Taqman specific primers and MGB probes for each serotype are designed and screened, and a real-time fluorescent PCR detection method is established. The detection system provided by the invention can be used for accurately, quickly and sensitively performing molecular biological detection on the three mimicry vibrios.

The invention discloses primers and a probe for real-time fluorescent quantitative PCR (polymerase chain reaction) detection of porcine epidemic diarrhea virus G2c subtype and application of the primers and the probe, and belongs to the field of animal virus molecular biology detection methods. By comparing S genes of different gene subtype strains of the porcine epidemic diarrhea virus, a pair of primers and a Taqman-MGB probe are designed on the basis of specific mutation sites of the G2c subtype strain, through virus solutions and clinical samples of the different gene subtype strains of the PEDV, it is proved that the primers can effectively distinguish G2c from other PEDV gene subtype strains, and the G2c and other PEDV gene subtype strains can be effectively distinguished. Meanwhile, no cross reaction exists on other viruses of the porcine intestinal diarrhea syndrome, and the detection sensitivity of the kit can reach 5 copies / microliter. According to the invention, the specific and sensitive detection of the G2c subtype can be realized, and the kit has positive significance on the diagnosis and epidemiological investigation of the porcine epidemic diarrhea virus.

The invention relates to the technical field of microbiological detection, and particularly discloses a composition and a kit for detecting vibrio harveyi, the composition for detecting vibrio harveyi is composed of a primer pair and a probe, the primer pair is composed of two single-stranded DNAs shown in a sequence 1 and a sequence 2 in a sequence table; the probe is a single-stranded DNA probe as shown in a sequence 3 in the sequence table (5'end is marked with a fluorophore FAM, and 3 'end is marked with a quenching group MGB). The method is high in sensitivity, accurate in result, good in repeatability, easy to operate and suitable for the fields of aquaculture and the like, a quenching group of the MGB probe is a non-fluorescent quenching group and does not generate fluorescence, the intensity of a background signal can be greatly reduced, the detection result is more accurate, and the purpose of rapidly and accurately detecting vibrio harveyi is achieved.